CN106755259A - A kind of pyrus bretschneideri rehd flesh cell method of counting - Google Patents
A kind of pyrus bretschneideri rehd flesh cell method of counting Download PDFInfo
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- CN106755259A CN106755259A CN201510826735.4A CN201510826735A CN106755259A CN 106755259 A CN106755259 A CN 106755259A CN 201510826735 A CN201510826735 A CN 201510826735A CN 106755259 A CN106755259 A CN 106755259A
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- kuerle delicious
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Abstract
A kind of pyrus bretschneideri rehd flesh cell method of counting of the present invention, with sarranine-fast green decoration method, the pyrus bretschneideri rehd section handled well to routine paraffin wax microtomy is dyeed, then the cell number in unit area is examined under a microscope, by based on the conversion method on the basis of fruit cross section area, calculate pyrus bretschneideri rehd flesh cell number, the inventive method can more accurately calculate pyrus bretschneideri rehd flesh cell number, it can be research pyrus bretschneideri rehd flesh cell volume, relation between flesh cell number and fruit size provides more scientific computational methods.
Description
Technical field
A kind of pyrus bretschneideri rehd flesh cell method of counting of invention, with sarranine-fast green decoration method, the pyrus bretschneideri rehd section handled well to routine paraffin wax microtomy is dyeed, then the cell number in unit area is examined under a microscope, by based on the conversion method on the basis of fruit cross section area, calculate pyrus bretschneideri rehd flesh cell number, this patent method can more accurately calculate pyrus bretschneideri rehd flesh cell number, it can be research pyrus bretschneideri rehd flesh cell volume, relation between flesh cell number and fruit size provides more scientific computational methods.
Background technology
Pears（Pyrus spp）Belong to the rose family（Rosaceae）Pears subfamily（Pomoideae）Pear（Pyrus.L）Plant, has more than 30 kind, is that the third-largest temperate zone fruit variety, there is the cultivation history of more than 3 000 years in the world after after grape and apple.Baile is divided into two classes according to the primary distribution of Pyrus L, and a class is west pears, or European pears（European pear）, also referred to as European pear originates from Mediterranean and Caucasus region, is distributed mainly on Europe, North America, South America, Africa and Oceania；Another kind of is east pears, also referred to as Asian pear（Asian pear）, originate from China, it is mainly distributed on the Asian countries such as China, Japan, South Korea.China is the first in the world Chan Li big countries, and the data display of statistics is announced according to USDA Foreign Agricultural Service：In 2012-2013 years, the yield of Chinese pear is 165 000 ten thousand tons, accounts for the 75% of world's pears total output, and 4 400 ten thousand tons of its middle outlet accounts for 25% or so of world's pears total volume of exports.Kuerle delicious pear is the famous characteristic fruit in Xinjiang, originates in Xinjiang Kurle region, is the natural hybrid species of pear and big desert pears, there is the cultivation history of more than 1 400 years.Kuerle delicious pear has thin skin meat crisp, and delicate succulence is fresh and sweet tasty and refreshing, and the features such as sugar content is high, Vitamin C content is abundant and storage tolerance, especially unique with fragrant odour.Once it is repeatedly prize-winning at home, the good reputations such as ' king in fruit ', ' treasure in pears ', ' China's honey pears ', ' after pears ', ' prince in pears ' are enjoyed in the international market, by development for many years, Kuerle delicious pear industry has become the pillar industry of local agro based economic development, it is also that peasant lifts oneself out of poverty, increase the main channel of income, the key farm products even more exported goods and earned foreign currency, domestic and international market portion is increasing, product finds a good sale in more than ten countries and regions such as Japan, Singapore, Thailand, Canada, deep to be liked by domestic and international consumer.
But Kuerle delicious pear has fruit less than normal, and pulp fraction is relatively small, the larger shortcoming of core part proportion.This characteristic fruit tree research center finds that the individual difference of size is caused by controlling the organ neural specific gene expression level to change between plant organ in early-stage Study.Pyrus bretschneideri rehd size is mainly what is together decided on by cell volume and cell number.After successfully pollinating, Fruit depends primarily on cell division and cell expands, and the two processes have together decided on the final volume of pyrus bretschneideri rehd.And Domestic Scientific Research worker is when Kuerle delicious pear pulp organization cell volume, number is studied, the method for being used is vertical and horizontal mensuration, error is larger, scientific research needs can not be met, the method for cell count on the basis of a kind of cross section area based on fruit has been invented in this experiment, can be used to different times Kuerle delicious pear pear flesh histocyte volume, number determine, for research Kuerle delicious pear flesh cell volume, the relation between cell number and pear fruit size provide more scientific computational methods.
The content of the invention
It is relatively small for pyrus bretschneideri rehd, edible pulp fraction is less, a kind of pyrus bretschneideri rehd flesh cell method of counting has been invented in the excessive feature in core part, the invention, in the research of the main cause that can be used for analyzing influence pyrus bretschneideri rehd size.This patent method has advantages below:1. the inventive method is the method for cell count carried out on the basis of pyrus bretschneideri rehd cross section area, more more scientific than traditional cell longitudinal and transverse demeter measuring method.2. inventive process avoids the troublesome operation step uneven, that be not readily separated cell of sampling of flow cytometry, with easy to operate, the characteristics of not strict to dyeing time requirement.3. the inventive method can more accurately position pulp confluent monolayer cells, and the interference of epidermal cell and vascular bundle cell is avoided to a greater extent, can be minimized the influence of lithocyte again.4. flesh cell can be dyed blueness by the inventive method, lithocyte and dredged histocyte and dyed black, with three-dimensional Color.5. the inventive method saves time and materials, it is not necessary to which slide can be observed directly, can also be fabricated to permanent section, for imparting knowledge to students, in scientific research.
Specific implementation method
1 cutting sample laterally cuts pyrus bretschneideri rehd along core, and it is r outwards to measure Kuerle delicious pear pears core partial radius successively from center with slide measure, and pyrus bretschneideri rehd radius is R.2 samples are fixed：Kuerle delicious pear pulp fraction is cut into the fritter of 0.5cm*0.5cm*1cm, is fixed in FAA fixers, change a fixer within the 2nd day, change a fixer within 4th day, a fixer is changed within 7th day, a fixer is changed within the 15th day, then by Sample storage in 4 ± 0.5 DEG C of refrigerators（It is 70% absolute alcohol that FAA fixes liquid making method:Glacial acetic acid:40% formalin=90:5:5）.3 reagents：Sarranine solution, it is standby after filtering during 1.2g sarranine dissolved in into the ethanol solutions of 100ml 50%；Fast green solution, dissolves in the ethanol solutions of 100ml 95% by 0.4g is fast green, standby after filtering.4 operating procedures：（1）Paraffin section makes and the Kuerle delicious pear pulp organization 30min that fixes → use 65% ethanol successively is embathed with 50% ethanol solution, 80% ethanol, 90% ethanol, 95% ethanol, 100% ethanol immersion 1.5h → be followed successively by 2: 1 in ethanol and dimethylbenzene volume ratio respectively, 1: 1, 2h → in 40 DEG C of baking ovens is soaked in 1: 2 mixed solution in each immersion 1h → pure dimethylbenzene, with dimethylbenzene and Quality of Paraffin Waxes ratio to soak the waxdip of 12 h → in 60 DEG C of baking ovens in 1: 1 mixed solution, wax liquor clean twice is changed daily, altogether change 6 times → Kuerle delicious pear pulp is cut into 10 μ m-thicks → paster → dewaxing → dyeing with routine paraffin wax microtomy（Sarranine Fast Green counterstain）→ neutral gum mounting.（2）Flesh cell number is calculated chooses Kuerle delicious pear pulp fraction under the microscope, counts certain area S（0.03cm2<S<0.05cm2）Interior flesh cell number N（The cell of unit area boundary is denoted as 1, fraction being denoted as 0) in unit area with the count method that rounds up, major part in unit area.The then average area S of individual cells（）=S/N, it is known that pyrus bretschneideri rehd radius is R, pears core partial radius are r, then surveyed Kuerle fruit fresh cell number is：
Claims (6)
1. a kind of pyrus bretschneideri rehd pulp fraction method for cell count, it is characterized in that first measuring pyrus bretschneideri rehd radius R with slide measure, Kuerle delicious pear pears nuclear radius r, Kuerle delicious pear pulp fraction sample is cut into after 0.5cm*0.5cm*1cm fritters and is immersed in FAA fixers, with the sarranine for preparing in advance, fast green dye liquor, the Kuerle delicious pear pulp fraction section handled well to routine paraffin wax microtomy is dyeed, through graded ethanol, after dimethylbenzene treatment, permanent section is fabricated to neutral gum, cell number under the microscope in observation unit area again, pyrus bretschneideri rehd pulp fraction cell number is calculated according to formula.
2. slide measure according to claim 1 measures pyrus bretschneideri rehd radius R, Kuerle delicious pear pears core partial radius r, it is characterized in that pyrus bretschneideri rehd is laterally cut along core, outwards measure Kuerle delicious pear pears nuclear radius r, pyrus bretschneideri rehd radius R successively from center.
3. sample according to claim 1 is fixed it is characterized in that Kuerle delicious pear pulp fraction is cut into 0.5cm*0.5cm*1cm fritters, it is fixed in FAA fixers, change a fixer within 2nd day, change a fixer within 4th day, change a fixer within 7th day, a fixer is changed within 15th day, then by Sample storage in 4 ± 0.5 DEG C of refrigerators.
4. the good Kuerle delicious pear pulp organization section of advanced processing according to claim 1, it is characterised in that：The Kuerle delicious pear pulp organization 30min that fixes → use 65% ethanol successively is embathed with 50% ethanol solution, 80% ethanol, 90% ethanol, 95% ethanol, 100% ethanol immersion 1.5h → be followed successively by 2: 1 in ethanol and dimethylbenzene volume ratio respectively, 1: 1, 2h → in 40 DEG C of baking ovens is soaked in 1: 2 mixed solution in each immersion 1h → pure dimethylbenzene, with dimethylbenzene and Quality of Paraffin Waxes ratio to soak the waxdip of 12 h → in 60 DEG C of baking ovens in 1: 1 mixed solution, wax liquor clean twice is changed daily, altogether change 6 times → Kuerle delicious pear pulp is cut into 10 μ m-thicks → paster → dewaxing → dyeing with routine paraffin wax microtomy（Sarranine Fast Green counterstain）→ neutral gum mounting.
5. the cell number in unit area according to claim 1, it is characterised in that：Kuerle delicious pear pulp fraction is chosen under the microscope, counts certain area S（0.03cm2<S<0.05cm2）Interior flesh cell number N（The cell of unit area boundary is denoted as 1, fraction being denoted as 0) in unit area with the count method that rounds up, major part in unit area.
It is 6. according to claim 1 that pyrus bretschneideri rehd pulp fraction cell number is calculated according to formula, it is characterised in that：
。
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| Application Number | Priority Date | Filing Date | Title |
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| CN201510826735.4A CN106755259B (en) | 2015-11-25 | 2015-11-25 | Method for counting fruit pulp cells of Korla fragrant pear |
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| CN201510826735.4A CN106755259B (en) | 2015-11-25 | 2015-11-25 | Method for counting fruit pulp cells of Korla fragrant pear |
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| CN106755259A true CN106755259A (en) | 2017-05-31 |
| CN106755259B CN106755259B (en) | 2021-05-25 |
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Cited By (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN113030088A (en) * | 2021-03-10 | 2021-06-25 | 浙江大学 | Cell rapid statistical evaluation method for solid lignification degree of loquat fruit |
Citations (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN101893469A (en) * | 2010-07-19 | 2010-11-24 | 南京农业大学 | Method for measuring volume of fruits |
| CN102102120A (en) * | 2010-12-20 | 2011-06-22 | 南京农业大学 | Method for fast measuring size and shape of fruit cells of pear |
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- 2015-11-25 CN CN201510826735.4A patent/CN106755259B/en active Active
Patent Citations (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN101893469A (en) * | 2010-07-19 | 2010-11-24 | 南京农业大学 | Method for measuring volume of fruits |
| CN102102120A (en) * | 2010-12-20 | 2011-06-22 | 南京农业大学 | Method for fast measuring size and shape of fruit cells of pear |
Non-Patent Citations (2)
| Title |
|---|
| 张振铭: "套袋对梨果实品质、石细胞及相关酶活性变化的影响", 《中国优秀硕士学位论文全文库 农业科技辑》 * |
| 张祖建等: "水稻胚乳细胞计数方法研究", 《江苏农学院学报》 * |
Cited By (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN113030088A (en) * | 2021-03-10 | 2021-06-25 | 浙江大学 | Cell rapid statistical evaluation method for solid lignification degree of loquat fruit |
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